CN1995319A - Multiple passage intelligent temperature-control device facing PCR microfluidic chip - Google Patents
Multiple passage intelligent temperature-control device facing PCR microfluidic chip Download PDFInfo
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- CN1995319A CN1995319A CN 200710062646 CN200710062646A CN1995319A CN 1995319 A CN1995319 A CN 1995319A CN 200710062646 CN200710062646 CN 200710062646 CN 200710062646 A CN200710062646 A CN 200710062646A CN 1995319 A CN1995319 A CN 1995319A
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Abstract
The invention discloses a multi-channel intelligent temperature-controlling device of polyase chain reaction micro-flow control chip, which comprises the following parts: three temperature sensors (3), three electric heat films (4), chip fixing device (1) and temperature collecting control device (2), wherein the chip fixing device is composed of three insulating areas (6), insulating groove (7) and pin on the insulating area with PCR micro-flow chip (5) on the surface of chip fixing device; the temperature collecting control device contains multi-path analog switch, A/D switcher, single chip set, D/A switcher, driving circuit and display.
Description
Technical field
The multiple passage intelligent temperature device of (PCR) micro-fluidic chip belongs to towards the microminiaturized integrated automation field of biochip towards the polymerase chain reaction.This device is mainly used in three flat-temperature zones that on the PCR micro-fluidic chip DNA increased and carries out respectively that independence is provided with and temperature control, total system require heat-up time short, volume is little, cost is low and can communicate with PC.
Background technology
The PCR micro-fluidic chip is a kind ofly novel can carry out the technology of amplification in vitro rapidly and efficiently to the dna fragmentation of multiple different lengths.Its principle be exactly with the PCR reaction mixture under the effect of precise injection pump, be injected into the microchannel that is in three flat-temperature zones on the substrate respectively with the flow velocity of nl/s, by the length of control microchannel, thereby realize sex change, renaturation and the extension of PCR.Its structure generally is made up of inlet, outlet, three flat-temperature zones, microchannel and heat insulation grids.Therefore be to influence the DNA successful key factor that can increase for the design of its three flat-temperature zone temperature controlling of PCR micro-fluidic chip, heat insulation grid.
The device that PCR micro-fluidic chip temperature control is at present generally adopted is: 1. copper billet heating; 2. temperature sensor measurement temperature; 3. miniature temperature controller controlled temperature.The shortcoming of this device is: the copper billet heating efficiency is low, total system is bulky, be unfavorable for detecting with integrated at sheet of chip.
Another device that the control of PCR micro-fluidic chip temperature is adopted is: 1. substrate (silicon/glass) lower surface deposit film heating; 2. substrate lower surface depositing temperature sensor measurement temperature simultaneously; 3. by adopting corresponding temperature acquisition control circuit to realize temperature control.The shortcoming of this device is: the heating film, and temperature sensor, the PCR micro-fluidic chip integrates, the cost height, the bonding difficulty of chip is big.
Summary of the invention
The objective of the invention is to solve the problem that present PCR micro-fluidic chip temperature control system exists, exploitation one cover heating efficiency height, volume are little, heating, temperature acquisition separate integrated with the PCR micro-fluidic chip, not only having reduced cost and also be beneficial in the sheet detection, is that a cover can carry out the independent integrated temperature control system of height that is provided with and controls to the temperature of three flat-temperature zones.The present invention has designed a cover multiple passage intelligent temperature device based on above-mentioned purpose, and this device comprises two portions: heating unit and temperature acquisition control device 2.Heating unit is referring to Fig. 3.The logical diagram of temperature acquisition control device 2 as shown in Figure 4.
Heating unit is made up of chip fixture apparatus 1, three temperature sensors 3, three Electric radiant Heating Film 4.Three temperature sensors 3, three Electric radiant Heating Film 4 are bonded in three flat-temperature zones of chip fixture apparatus 1 lower surface respectively by thermostable heat-conductive glue, wherein three Electric radiant Heating Film 4 link to each other with second pin 17 respectively, three temperature sensors 3 link to each other with first pin 8 respectively, chip fixture apparatus 1 is connected with temperature acquisition control device 2 by lead, and first pin 8 is connected with temperature acquisition control device 2 by lead with second pin 17.PCR micro-fluidic chip 5 is bonded in the upper surface of chip fixture apparatus 1 by heat-conducting glue in addition.Wherein PCR micro-fluidic chip 5 comprises: three flat-temperature zones 9, insulation hole 10, microchannel 11, inlet 12 and export 13, the flat-temperature zone 9 of PCR micro-fluidic chip 5, insulation hole 10 flat-temperature zone 6, the heat dam 7 with chip fixture apparatus 1 respectively are corresponding respectively.It is heat insulation to adopt the air heat dam to carry out between three flat-temperature zones on the chip fixture apparatus 1 in addition, and the width of groove is not less than 4mm, and chip fixture apparatus 1 adopts metallic substance, and material thickness is not less than 1mm, can adopt the copper of good heat conductivity.It is heat insulation to adopt the insulation hole that is not less than 2mm to carry out above the PCR micro-fluidic chip 5, and PCR micro-fluidic chip 5 selects high polymer material as substrate.Wherein the vertical view of PCR micro-fluidic chip 5 as shown in Figure 3.
Temperature acquisition control device 2 is characterised in that: be made up of multiway analog switch, A/D converter, micro-chip, D/A converter, driving circuit and indicating meter.The signal that No. three temperature sensors are gathered is through amplifying circuit, deliver on the multiway analog switch, multiway analog switch selects the signal on a certain road to give A/D converter, A/D converter sends numerary signal to micro-chip, micro-chip compares the signal that collects and the temperature that is provided with on this road, if acquired signal is less than temperature is set, then micro-chip output control signal is given the D/A chip, the D/A chip is then exported the voltage of setting and is given driving circuit, driving circuit is exported the Electric radiant Heating Film required voltage, and it is heated.Otherwise if greater than temperature is set, then micro-chip output control signal is given the D/A chip, and the D/A chip is then exported the voltage of setting and given driving circuit, stops the heating to Electric radiant Heating Film.Micro-chip is seen Fig. 5 to the logical diagram of temperature acquisition control device, and temperature acquisition control device main program flow chart and temperature acquisition control device judgement jump subroutine schema are seen Fig. 6 and Fig. 7 respectively.
Advantage behind the employing said apparatus is:
1) volume of temperature control unit reduces, and it is big to be reduced to palm by the body of original desktop computer size.
2) temperature control unit can carry out independent setting, control to three road warm areas simultaneously and show in real time.
3) experimental results show that this device can play effect of heat insulation, can satisfy the requirement of DNA being carried out the thermograde of external amplification.
Description of drawings:
Fig. 1: chip fixture apparatus vertical view
Among the figure: 6, flat-temperature zone, 7, heat dam, 8, first pin, 17, second pin
Fig. 2: PCR micro-fluidic chip vertical view
Among the figure: 9, flat-temperature zone, 10, insulation hole, 11, the microchannel, 12, inlet, 13, outlet
Fig. 3: chip fixture apparatus sectional view
Among the figure: 1, chip fixture apparatus, 3, temperature sensor, 4, Electric radiant Heating Film 5, PCR micro-fluidic chip, 11, the microchannel on the PCR micro-fluidic chip
Fig. 4: temperature acquisition control device schematic diagram
Among the figure: 2, temperature acquisition control device
Fig. 5: temperature acquisition control device logical diagram
Fig. 6: temperature acquisition control device main program flow chart
Fig. 7: the temperature acquisition control device system is judged the jump subroutine schema
Fig. 8: the chip fixture apparatus heat dam is counted temperature field value mimic diagram
Among the figure: 14, inlet, 15, outlet, 16, tube wall
Fig. 9: chip fixture apparatus heat dam thermal infrared imager figure
Embodiment:
Chip fixture apparatus the present invention towards the multiple passage intelligent temperature device of PCR micro-fluidic chip uses the good copper sheet of heat conductivility as body material.This chip fixture apparatus adopts the air band between the heat dam of wide 4mm, long 60mm to carry out heat insulation.The present invention detects three aspects from Theoretical Calculation, numerical simulation, experiment and proves the feasibility of this device.
Theoretical Calculation: the heat transfer problem of heat dam mainly divides three parts: the 1. adjacent copper sheet of heat dam thermal radiation separately
2. the radiation heat transfer q between the adjacent copper sheet of heat dam
1,2=q
2,1=X
2,1(q
R1-q
R2); 3. the convective heat exchange q of air between the adjacent copper sheet of heat dam
Air=α Δ T.Wherein ε is a blackness, is 0.22 in the present embodiment; σ is blackbody radiation constant 5.67w/ (m
2K
4); The temperature of object when T is thermal equilibrium; X
1,2Being two angle factors between the face, is 0.4 in the present embodiment.α is the convection transfer rate of air, is 6 in the present embodiment; The temperature of air is 25 ℃.According to the physical parameter of above-mentioned formula in conjunction with each material, the heat transfer situation that can calculate between the adjacent copper sheet of heat dam sees Table 1.In the table 1: for heat dam 1, T
W1And T
W2Be the temperature on included adjacent two the copper sheet surfaces of heat dam 1, be respectively 94 ℃ and 72 ℃, the thermal radiation on two surfaces is respectively q
1(229.8w/m
2) and q
2(179.5w/m
2).For heat dam 2, T
W1And T
W2Be the temperature on included adjacent two the copper sheet surfaces of heat dam 2, be respectively 72 ℃ and 55 ℃.The thermal radiation on two surfaces is respectively q
1(179.5w/m
2) and q
2(142.6w/m
2).Then as can be seen from Table 1, heat dam 1 and 2 net radiation heat flow density (q
1+ q
2-2q
1,2) be respectively 368.2w/m
2And 292.2w/m
2, and the heat flow density that the air of heat dam 1 and 2 narrow spaces is taken away because of natural convection is respectively: 696w/m
2And 462w/m
2, so the air band between width 4mm, long 60mm heat dam can play heat insulation effect.
Heat transfer between the adjacent copper sheet of table 1
| T w1 | T w2 | q 1 | q 2 | 2q 1，2 | q 1+q 2-2q 1，2 | q air |
| Heat dam | (℃) | (℃) | (w/m 2) | (w/m 2) | (w/m 2) | (w/m 2) | (w/m 2) |
| 1 2 | 94 72 | 72 55 | 229.8 179.5 | 179.5 142.6 | 41.1 29.8 | 368.2 292.2 | 696 462 |
Numerical simulation: use GAMBIT and created long 60mm, wide 4mm, the model of dark 2mm, model is made up of inlet 14, outlet 15 and tube wall 16.After application FLUENT6.1 reads this project, it is as follows that final condition is set: pipe wall material copper, the temperature in tube wall left side is set to 345K, the temperature on right side is set to 367K, and working fluid is an air, adopts natural convection air to carry out heat insulation, gas inlet place temperature is 300K, when the speed of natural convection air is generally less than 0.3m/s, select 0.03m/s in the present embodiment, the direction of natural convection perpendicular to horizontal direction along Z axially down.Analog result is seen Fig. 9.Working as speed as can be seen by this figure is 0.03m/s, temperature be the air of 300K through long 60mm, wide 4mm is behind the heat insulating belt of dark 2mm, utilize the natural convection of air can take away the heat that is produced by the thermal radiation of two constant temperature faces, promptly this heat insulating belt can play heat insulation effect.For same engineering, under the constant condition of other simulated conditions, change the right flank temperature value into 328K, then the numerical simulation result that distributes of the temperature field of 345K and the adjacent warm area of 328K is consistent with Fig. 9.The results of numerical simulation represents that the heat dam that present embodiment adopts can satisfy the test requirements document of carrying out the dna amplification reaction thermograde.
Experimental verification: for the effect of heat insulation of the heat dam of wide 4mm, long 60mm, present embodiment is used the NECTH5102 infrared thermography and has been measured temperature field on the whole copper sheet and distribute and see Figure 10.Temperature field on the copper sheet is divided significantly for three flat-temperature zones as seen from Figure 10, and the medial temperature of three flat-temperature zones is respectively: 94.3 ℃, 72.1 ℃ and 55.6 ℃; Thermograde between adjacent warm area is 22.2 ℃ and 16.5 ℃.This explanation Theoretical Calculation, numerical simulation result are correct.The wide 4mm that is adopted in promptly inventing, the heat dam of long 60mm can play heat insulation effect, can satisfy the requirement to thermograde of the external amplification of DNA.
The temperature regulating device of adorning towards the multiple passage intelligent temperature of PCR micro-fluidic chip is: adopt 3 road AD590 as temperature sensor, the signal of its collection amplifies through amplifier and is converted to voltage signal, three road voltage signals are connected with multiway analog switch, select one road voltage signal through the MCS-2051 micro-chip control, convert this signal to numerary signal through the A/D conversion chip, the temperature value of this signal and setting compares, if give the D/A conversion chip less than temperature MCS-2051 micro-chip output control signal is set, D/A chip output " 5V " voltage is given driving circuit, driving circuit amplifies input voltage, exports to Electric radiant Heating Film and heats; If instead greater than the temperature that is provided with, then MCS-2051 micro-chip output control signal is given the D/A conversion chip, and driving circuit is given in D/A chip output " 0v ", and Electric radiant Heating Film is stopped heating.This device is weaved into and is realized by 2051 micro-chips being carried out assembly language the collection of temperature and control, and wherein temperature acquisition control device main program flow chart and temperature acquisition control device judge that the jump subroutine schema sees Fig. 6 and Fig. 7 respectively.
Temperature sensor signal is an electric quantity signal with non-electro-temperature conversion of signals in the temperature regulating device, and electric quantity signal connects amplifier reverse input end 2, positive input 3 ground connection, and 6 is output terminal.The adjusting of sample circuit comprises zeroing and transfers full.Wherein zeroing is meant: regulate slide rheostat R3, make the voltage that is added on the resistance R (VR3+R5) be+5V, the electric current that the electric current of its generation and temperature sensor are exported in the time of 0 ℃ offsets, and makes that the input terminus 2 of operational amplifier is zero, and then output terminal 6 is zero volt.Accent is full of finger: in the time of 100 ℃, regulate slide rheostat R4, make that output voltage is 5V.In order to obtain stable magnitude of voltage, resistance R answers the little precision resistance of use temperature coefficient and use+5v voltage stabilizing tube to stablize input voltage.
The voltage signal that obtains from No. three sample circuits meets the X0 of multiway analog switch respectively, X1, the X2 port, the A of multiway analog switch, B control port meet the P1.6 and the P1.7 port of MCS-51 series monolithic, pin X3～X7, INH, GND, VEE, C ground connection, VDD connects+the 12V power supply, and 3 meet the input pin CH1 of A/D conversion chip.The CH0 of A/D conversion chip and DI and GND ground connection, VCC connects+power supply of 5V, CH1 connects input signal, and DO, CLK, CS meet P1.5, P1.4 and the P1.3 of 2051 chips respectively, with convenient 2051 chips it are controlled.
The pin mode of connection of MCS-51 series monolithic is: VCC connects+and the working power of 5V, GND ground connection, RST is as the reset signal input terminus, the outside by 10 μ F electric capacity and+power supply of 5V joins, P1.7, the A of P1.6 and multiway analog switch links to each other with B control input (output) end, P1.5, P1.4 and P1.3 respectively with the DO of TLC0832 type A/D conversion chip, CLK, CS divides and joins, XTAL1, XTAL2 and clock generator join, external crystal oscillator frequency is 11.0592MHz, P3.2, P3.3, the display port of P3.7 and liquid-crystal display joins, P1.2, P3.4, P3.5 respectively with the LOAD of D/A conversion chip, CLK, the DATA port joins.Other 2051 micro-chips can join by ROUT, the TIN port of P3.0, P3.1 and serial communication chip MAX232, thereby realize and the communicating by letter of PC.
The mode of connection of D/A conversion chip pin is: pin (LOAD) and pin (LDAC) are the control ends of output, pin (LDAC) ground connection, make its function out of hand, the output signal of D/A chip is only controlled the negative edge output data by pin (LOAD) like this.The channel selecting of D/A conversion chip is by the 4th and the 5th control of serial input signals.The reference voltage of DS5620 chip all is set to+5V, and the formula of its D/A conversion is seen (1):
V＝REF*CODE*(1+RNG)/256 (1)
Wherein CODE is the numerary signal of 2051 micro-chips output, and the value of establishing RNG is 1, and when the digital quantity of input reached 80H (ten advance value 128), the analog quantity of output was+5V like this.When the digital quantity of input during greater than 80H, the analog quantity of output is maximum value+5V.
The voltage of D/A chip output is through by two magnifications that integrated operational amplifier constituted being 2.4 times forward voltage amplifying circuit, and the common-collector amplifier of being made up of middle power NPN type triode provides electric current to amplify to export then.Design of drive circuit depends on the rated operational voltage and the electric current of Electric radiant Heating Film.
The indicating meter of this device is connected with the MCS-51 series monolithic by the socket of one 8 stature.Its mode of connection in system is as follows: pin 1VSS connects power supply ground, 2VCC connects+power supply of 5V, 3VEE connects-power supply of 12V by slide rheostat, 4CLK connects the P3.2 pin of MCS-51 series monolithic, 5SID connects the P3.3 pin of 2MCS-51 series monolithic, the 6CS sheet selects the P3.7 pin of termination MCS-51 series monolithic, and 7 backlight voltages connect+5V, 8 backlight common ports.
Claims (5)
1, towards the multiple passage intelligent temperature device of polymerase chain reaction micro-fluidic chip, it is characterized in that: the chip fixture apparatus (1) and the temperature acquisition control device (2) that comprise three temperature sensors (3), three Electric radiant Heating Film (4), installation three temperature sensors (3), three Electric radiant Heating Film (4), PCR micro-fluidic chip (5); Wherein, chip fixture apparatus (1) is made up of the heat dam (7) between three flat-temperature zones (6), the flat-temperature zone and first pin (8) on the flat-temperature zone and second pin (17), the chip fixture apparatus upper surface is installed PCR micro-fluidic chip (5), and flat-temperature zone (6), heat dam (7) are corresponding respectively with flat-temperature zone (9), the insulation hole (10) of PCR micro-fluidic chip (5); Three Electric radiant Heating Film (4) and three temperature sensors (3) are separately positioned on three flat-temperature zones of chip fixture apparatus (1) lower surface, wherein three Electric radiant Heating Film (4) link to each other with second pin (17) respectively, three temperature sensors (3) link to each other with first pin (8) respectively, and chip fixture apparatus (1) is connected with temperature acquisition control device (2) by lead.
2, the multiple passage intelligent temperature device towards the polymerase chain reaction micro-fluidic chip according to claim 1, it is characterized in that: chip fixture apparatus (1) adopts metallic substance, material thickness is more than or equal to 1mm, and the width of heat dam (7) is more than or equal to 4mm.
3, the multiple passage intelligent temperature device towards the polymerase chain reaction micro-fluidic chip according to claim 2, it is characterized in that: described metallic substance is a copper.
4, the multiple passage intelligent temperature device towards the polymerase chain reaction micro-fluidic chip according to claim 1 is characterized in that: three Electric radiant Heating Film (4) stick on three flat-temperature zones of chip fixture apparatus (1) lower surface respectively by thermostable heat-conductive glue.
5, multiple passage intelligent temperature device towards the polymerase chain reaction micro-fluidic chip according to claim 1, it is characterized in that: described temperature acquisition control device (2) is mainly by multiway analog switch, A/D converter, micro-chip, D/A converter, driving circuit and indicating meter are formed: the signal that No. three temperature sensors are gathered is through amplifying circuit, deliver on the multiway analog switch, multiway analog switch is used to select the signal on a certain road to give A/D converter, A/D converter sends numerary signal to micro-chip, micro-chip compares the signal that collects and the temperature that is provided with on this road, and D/A converter is used for control signal with micro-chip output and is converted to voltage signal and exports to driving circuit; When the micro-chip acquired signal less than temperature is set, D/A converter is then exported the voltage of setting and is given driving circuit, driving circuit output Electric radiant Heating Film required voltage, and it is heated; When acquired signal greater than temperature is set, D/A converter is then exported the voltage of another setting and is given driving circuit, makes it stop heating to Electric radiant Heating Film; Indicating meter links to each other with micro-chip, is used to show the temperature of certain warm area of real-time collection.
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|---|---|---|---|
| CN 200710062646 CN1995319A (en) | 2007-01-12 | 2007-01-12 | Multiple passage intelligent temperature-control device facing PCR microfluidic chip |
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|---|---|---|---|
| CN 200710062646 CN1995319A (en) | 2007-01-12 | 2007-01-12 | Multiple passage intelligent temperature-control device facing PCR microfluidic chip |
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| CN102304464A (en) * | 2011-10-08 | 2012-01-04 | 武汉馨世生物科技有限公司 | Multiplex polymerase chain reaction amplification instrument |
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| CN102791847A (en) * | 2010-01-12 | 2012-11-21 | 阿赫姆生物系统公司 | Three-stage thermal convection apparatus and uses thereof |
| CN103194383A (en) * | 2013-04-09 | 2013-07-10 | 湖北民族学院 | Chip level PCR-LVCE integrated system |
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| CN102304464A (en) * | 2011-10-08 | 2012-01-04 | 武汉馨世生物科技有限公司 | Multiplex polymerase chain reaction amplification instrument |
| CN103194383B (en) * | 2013-04-09 | 2014-09-03 | 湖北民族学院 | Chip level PCR-LVCE integrated system |
| CN103194383A (en) * | 2013-04-09 | 2013-07-10 | 湖北民族学院 | Chip level PCR-LVCE integrated system |
| CN103215184B (en) * | 2013-04-12 | 2014-07-30 | 北京理工大学 | Micro gene amplification device for miniature space experiment |
| CN103215184A (en) * | 2013-04-12 | 2013-07-24 | 北京理工大学 | Micro gene amplification device for miniature space experiment |
| CN105316226A (en) * | 2014-05-31 | 2016-02-10 | 捷普科技（上海）有限公司 | Lifting mechanism and biochip detection device with same |
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